Electrospray ionisation mass spectrometry facilitates detection of fibrinogen (Bbeta 14 Arg --> Cys) mutation in a family with thrombosis.

Abstract

We report the first direct detection of a fibrinogen mutation by electrospray ionisation mass spectrometry. The propositus, from a family with a history of thrombosis, came to attention after a pulmonary embolism subsequent to a spontaneous abortion. Prolonged thrombin (41 s) and reptilase times (26 s) together with an impairment of fibrinopeptide B release suggested a mutation at the thrombin cleavage site of the Bbeta chain. Direct mass analysis of purified fibrin chains from a thrombin induced clot showed that 50% of the Bbeta chains remained uncleaved. The measured mass of the mono sialo isoform of this uncleaved chain was 54150 Da, compared to a value of 54198 Da for normal Bbeta chains. This decrease of 48 Da in the intact protein is indicative of either a Bbeta 14 Arg to Cys, or Arg to Leu substitution. Heterozygosity for the Bbeta 14 Arg --> Cys mutation was verified by PCR amplification and DNA sequence analysis.

• Authors: S O Brennan, B Hammonds, R Spearing, P M George
• Journal: Thrombosis and haemostasis (Thromb Haemost) Vol. 78 Issue 6 Pg. 1484-7 (Dec 1997) ISSN: 0340-6245 [Print] Germany
• PMID: 9423799 (Publication Type: Case Reports, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Glycoproteins
• Fibrinopeptide A
• Fibrinopeptide B
• Fibrin
• Fibrinogen
• Arginine
• Thrombin
• Cysteine

Topics

• Adult
• Amino Acid Substitution
• Arginine (genetics)
• Catalysis
• Cysteine (genetics)
• Family Health
• Female
• Fibrin (analysis, chemistry)
• Fibrinogen (analysis, chemistry)
• Fibrinopeptide A (metabolism)
• Fibrinopeptide B (analysis, chemistry, genetics)
• Glycoproteins (analysis)
• Heterozygote
• Humans
• Mass Spectrometry (methods)
• Molecular Weight
• Mutation (genetics, physiology)
• Point Mutation (genetics, physiology)
• Polymerase Chain Reaction
• Sequence Analysis
• Thrombin (chemistry)
• Thrombosis (genetics)